Azole compounds endowed with antimycotic activity for human and veterinary use

ABSTRACT

Compounds of formula ##STR1## wherein R 1  is chloro, fluoro, bromo or trifluoromethyl; 
     R 2  is hydrogen, chloro , fluoro, bromo or trifluoromethyl; 
     Z is CH or N; 
     R 3 , R 4  and R 5 , which are the same or different, are hydrogen or C 1  -C 4  alkyl, with the proviso that R 4  is different from R 5  when R 3  is hydrogen; 
     X is O, S, SO or SO 2  ; 
     R 6  is a C 1  -C 5  polyfluoroalkyl group containing at least two fluorine atoms and optionally other halogen atoms selected from the group consisting of chloro and bromo; processes for their preparation and pharmaceutical compositions containing them are described. 
     The compounds of formula II-A are endowed with a marked wide range antimycotic acitivity.

This application is a Divisional of application Ser. No. 08/806,149,filed Feb. 25, 1997. Now U.S. Pat. No. 5,869,512.

The present invention relates to antimycotic compounds for human andveterinary use, and particularly, relates to azole compounds endowedwith antimycotic activity for the treatment and prophylaxis ofinfections in human and animals due to fungi and yeasts.

Among the antimycotic compounds known in literature an important classis composed by the so-called azole derivatives which embraces somecompounds employed in therapy such as Fluconazole (The Merck Index, XIed., No.4054, page 645), Itraconazole (The Merck Index, XI ed., No.5131, page 825) and Ketoconazole (The Merck Index, XI ed., No. 5181,page 835).

However, as far as we know, no one of these compounds is endowed with amarked antimycotic activity against some opportunistic pathogenic fungalstrains which cause infections even fatal for immuno-depressed patients.

Among the azole derivatives known as antimycotic for human or veterinaryuse a number of compounds characterised by the presence of a tertiaryalcohol moiety in their formula ##STR2## wherein Az represents atriazole or imidazole group, X' is preferably chloro, fluoro ortrifluoromethyl, R is preferably hydrogen, chloro or fluoro, R' and R",which are the same or different, are hydrogen or alkyl groups, Y is S,SO, SO₂ or O, and A is alkyl; hereinafter named as tertiary alcoholazoles, are described.

Among these tertiary alcohol azoles we mention, for example, thecompounds described in the European patent applications No. 54974(Sumitomo Chemical Company Limited), No. 61835 (Imperial ChemicalIndustries PLC), No. 107392 (Pfizer Limited), No. 140154 (SumitomoChemical Company Limited), No. 178533 (Sumitomo Pharmaceutical CompanyLimited), No. 435081 (SS Pharmaceutical Co. Ltd.) and No. 473387 (SankyoCompany Limited).

For some of these compounds a marked antimycotic activity was disclosed,though generically sometimes, both topically and systemically. However,as far as we know, the only compound under development is the one knownas Genaconazole, (2R,3R)α-(2,4-difluorophenyl)-α-[1-(methylsulfonyl)ethyl]-1H-1,2,4-triazol-1-ethanol,disclosed in the European patent application No. 178533.

Recently higher homologues of the tertiary alcohol azoles of formula Ihave been described in the European patent application No. 679647 (NihonNohyaku Co. Ltd.) as endowed with a good antimycotic activity againstCandida albicans, and characterised in that A is an optionallysubstituted phenyl or heterocycle.

The International patent application No. WO 96/31490 in the name of thesame applicants discloses the antimycotic activity of racemic mixturesof some azole compounds of formula II.

It has been now found that single enantiomers of the azole derivativesof formula ##STR3## wherein R₁ is chloro, fluoro, bromo ortrifluoromethyl;

R₂ is hydrogen, chloro, fluoro, bromo or trifluoromethyl;

Z is CH or N;

R₃, R₄ and R₅, which are the same or different, are hydrogen or C₁ -C₄alkyl, with the proviso that R₄ is different from R₅ when R₃ ishydrogen;

X is O, S, SO or SO₂ ;

R₆ is a C₁ -C₅ polyfluoroalkyl group containing at least two fluorineatoms and optionally other halogen atoms selected from the groupconsisting of chloro and bromo;

are endowed with a marked wide range antimycotic activity against fungipathogenic for human and animals, particularly also against fungalstrains resistant to antimycotics used in therapy, and againstopportunistic pathogenic fungal strains causing infections inimmuno-depressed subjects, and they are active both topically andsystemically. Some of the compounds of formula II in racemic mixture,especially the ones wherein X is O or S, are comprised in the Europeanpatent application No. 315946 (Presidenza del Consiglio deiMinistri--Ufficio del Ministero per il Coordinamento delle Iniziativeper la Ricerca Scientifica e Tecnologica) and said to be useful inagriculture as immunising agents against fungal pathologies and asphytogrowth regulators for useful growings.

Therefore object of the present invention are compounds of formula##STR4## wherein R₁ is chloro, fluoro, bromo or trifluoromethyl;

R₂ is hydrogen, chloro, fluoro, bromo or trifluoromethyl;

Z is CH o N;

R₃, R₄ and R₅, which are the same or different, are hydrogen or C₁ -C₄alkyl, with the proviso that R₄ is different from R₅ when R₃ ishydrogen;

X is O, S, SO or SO₂ ;

R₆ is a C₁ -C₅ polyfluoroalkyl group containing at least two fluorineatoms and optionally other halogen atoms selected from the groupconsisting of chloro and bromo;

and the pharmaceutically acceptable acid salts thereof.

The proviso at formula II-A is made to exclude the compounds claimed inthe European patent application No. 272679.

Another object of the present invention is constituted by compounds offormula II-A and the pharmaceutically acceptable salts thereof for useas a medicament.

The carbon atom marked with an asterisk has the absolute configurationdefined in the formula. Thus according to the Cahn, Ingold and Prelogconvention, the configuration of this carbon atom is R or S depending onthe priority order of the substituents.

The compounds of formula II-A can also contain a second chiral centrewhen R₃ and R₄ are different one from the other, and thus bediastereoisomers. The diastereoisomers of formula II-A object of theinvention are threo diastereoisomers.

The compounds of formula II-A are endowed with a powerful wide rangeantimycotic activity, particularly against Candida spp and Cryptococcusneoformans strains resistant to Fluconazole and Itraconazole, andagainst Candida glabrata, Candida krusei, Aspergillus spp and Fusariumspp resistant to Itraconazole and, like the previous ones, againstpathogenic strains responsible of fungal infections in immuno-depressedpatients, and are useful for the treatment and prophylaxis of fungal andyeast infections in human and animals.

The term C₁ -C₄ alkyl for R₃, R₄ and R₅ means methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl and t.butyl groups, methyl andethyl being preferred.

The term C₁ -C₅ polyfluoroalkyl group containing at least two fluorineatoms preferably intends difluoromethyl, trifluoromethyl,1,1,2-trifluoro-2-chloroethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl,1,1,2-trifluoroethyl, 1,1,2,2-tetrafluoroethyl,1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3-tetrafluoropropyl groups and theisomers thereof, the 1,1,2,2-tetrafluoroethyl group being the mostpreferred.

The salts of the compounds of formula II-A are salts withpharmaceutically acceptable organic and inorganic acids, such ashydrochloric, hydrobromic, hydroiodic, nitric, phosphoric, acetic,oxalic, malic, benzoic, benzenesulphonic, methansulphonic,4-methyl-benzenesulphonic, fumaric, lactic, tartaric, citric andgluconic acid. Preferred compounds of formula II-A are the ones whereinR₁ is chloro or fluoro, R₂ is hydrogen, chloro or fluoro, R₃ is methylor ethyl, R₄ and R₅, which are the same or different, are hydrogen,methyl or ethyl, Z is N and R₆ is a 1,1,2,2-tetrafluoroethyl group.

More preferred compounds of formula II-A are the ones wherein R₁ ischloro or fluoro, R₂ is hydrogen, chloro or fluoro, R₃ is methyl orethyl, R₄ and R₅, which are the same or different are hydrogen, methylor ethyl, Z is N, R₆ is a 1,1,2,2-tetrafluoroethyl group and X is O orSO₂.

Specific examples of the preferred compounds of formula II-A are thefollowing:

(2R,3S)1-(1H-1,2,4-triazolyl)-2-(2,4-difluorophenyl)-3-ethyl-4-(1,1,2,2,-tetrafluoro-ethoxy)-2-butanol

(2R,3S)1-(1H-1,2,4-triazolyl)-2-(2,4-dichlorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoro-ethoxy)-2-butanol

(2R,3R)1-(1H-1,2,4-triazolyl)-2-(2,4-dichlorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoro-ethylthio)-2-butanol

(2R,3S)1-(1H-1,2,4-triazolyl)-2-(2,4-difluorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoro-ethoxy)-2-butanol

(2R,3R)1-(1H-1,2,4-triazolyl)-2-(2,4-difluorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoro-ethylthio)-2-butanol

(2R,3S)1-(1H-1,2,4-triazolyl)-2-(4-chlorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoro-ethoxy)-2-butanol

(2R,3R)1-(1H-1,2,4-triazolyl)-2-(4-chlorophenyl)-3-methyl-4-(1,1,2,2,-tetrafluoro-ethylthio)-2-butanol

(2R)1-(1H-1,2,4-triazolyl)-2-(2,4-difluorophenyl)-3,3-dimethyl-4(1,1,2,2,-tetrafluoro-ethoxy)-2-butanol

(2R,3S)1-(1H-1,2,4-triazolyl)-2-(4-fluorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoro-ethoxy)-2-butanol

(2R,3R)1-(1H-1,2,4-triazolyl)-2-(4-fluorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoro-ethylthio)2-butanol

(2S)1-(1H-1,2,4-triazolyl)-2-(4-chlorophenyl)-3,3-dimethyl-4-(1,1,2,2-tetrafluoro-ethoxy)-2-butanol

(2S)1-(1H-1,2,4-triazolyl)-2-(4-chlorophenyl)-3,3-dimethyl-4-(1,1,2,2-tetrafluoro-ethylthio)-2-butanol

(2S)1-(1H-1,2,4-triazolyl)-2-(4-fluorophenyl)-3,3-dimethyl-4-(1,1,2,2-tetrafluoro-ethoxy)-2-butanol

(2S)1-(1H-1,2,4-triazolyl)-2-(4-fluorophenyl)-3,3-dimethyl-4-(1,1,2,2-tetrafluoro-ethylthio)-2-butanol

(2R,3R)1-(1H-1,2,4-triazolyl)-2-(2,4-dichlorophenyl)-3-methyl-4-(1,1,2,2-tetrachloroethylsulfonyl)-2-butanol.

The preparation of the compounds of formula II-A wherein X is O or S maybe carried out according to the synthetic scheme described in thealready cited European patent application No. 315946 starting from thesuitable intermediates in optically active form.

The compounds of formula II-A wherein X is SO or SO₂ may be obtainedfrom the corresponding compounds of formula II-A wherein X is Sfollowing conventional oxidation techniques. Preferably hydrogenperoxide, hypohalides or peracids are employed as oxidising agents,optionally in the presence of catalysts. Preferably the preparation ofthe compounds of formula II-A wherein X is O or S is carried out byadding a polyfluoroolefin of formula ##STR5## wherein X₁ and X₂, whichare the same or different, are F, Cl or CF₃ ;

or by reacting a polyfluorinated alcohol of formula

    R.sub.7 --CH.sub.2 --OH                                    (IV)

wherein R₇ is a C₁ -C₄ polyfluoroalkyl group containing at least twofluorine atoms and optionally other halogen atoms selected from thegroup consisting of chloro and bromo;

with an optically active intermediate of formula ##STR6## wherein R₁,R₂, R₃, R₄, R₅ and Z are as defined above, and X is O or S;

or with a reactive derivative thereof such as an ester, for example, amesylate, tosylate or trifluoromethansulphonate.

The optically active intermediates of formula V may be preparedfollowing known methods.

For example, the compounds of formula V may be prepared by theresolution of the corresponding racemic mixtures such as the onesdescribed in the above cited European patent application No. 315946 orin the U.S. Pat. No. 5134152.

Alternatively, the compounds of formula V wherein X is S may be preparedfrom the corresponding compounds of formula V wherein X is O throughknown methods. Furthermore the enzymatic resolution by stereoselectiveacylation and subsequent hydrolysis was found particularly advantageousfor preparing the compounds of formula V wherein X is O and R₃ and R₄are both methyl.

Though the enzymatic resolution by stereoselective acylation is knownfor preparing the lower homologous tertiary alcohol of the compounds offormula V (see, for example, the International patent application WO94/24305--Zeneca Limited), is surprising that it may also be applied forpreparing the compounds of formula V wherein X is O and R₃ and R₄ areboth methyl.

In fact, not only in these compounds there are two carbon atoms betweenthe moiety to be acylated and the asymmetric carbon atom, but above allone of these carbon atoms is disubstituted thus bearing a conspicuoussteric hindrance.

Therefore a further object of the present invention is a process forenzymatically resolving the compounds of formula V wherein X is O, andR₃ and R₄ are both methyl, process comprising their stereoselectiveacylation and subsequent hydrolysis.

Alternatively, the compounds of formula V wherein X is O and R₅ is ahydrogen atom may be prepared by reducing the corresponding acids offormula ##STR7## wherein R₁, R₂, R₃, R₄ and Z are defined as above.

The compounds of formula VI are known or can be readily preparedaccording to known methods (Bartroli J. et al., J. Org. Chem. 1995, 60,3000-3012).

The preparation of the salts of the compounds of formula II-A may becarried out according to conventional techniques, for example, byadmixing in solution equimolar amounts of a compound of formula II-A andof the selected acid, and separating the salt by precipitation andfiltration or evaporation of the solvent.

The compounds of formula II-A and their salts are antimycotic compoundsuseful in the treatment and prophylaxis of fungal and yeast infectionsin human and animals. Actually, the compounds of formula II-A object ofthe present invention are endowed with antimycotic activity againstyeast, filamentosous fungi, dermatophytes and dimorphic fungi.

The antimycotic activity has been evaluated in vitro as IC₅₀ and as MIC(minimum inhibiting concentration) on a number of strains such as, forexample, Candida albicans, Cryptococcus neoformans, Trichophytonmentagrophytes, Aspergillus fumigatus, Candida parapsilosis, Candidalusitaniae, Candida kefyr, Candida tropicalis, Candida krusei, Candidaglabrata, Aspergillus niger and Fusarium spp.

It is important to underline that the compounds of formula II-A objectof the present invention have shown to be effective against all thestrains of Candida spp. and Cryptococcus neoformans taken intoconsideration, comprising the ones resistant to Fluconazole,Itraconazole and Genaconazole.

A particularly marked antimycotic activity was revealed also againststrains of Candida glabrata and Fusarium spp., resistant to Itraconazoleand Genaconazole, and against Candida krusei and Aspergillus fumigatus,resistant to Fluconazole and Genaconazole, all of them being pathogenicagents responsible for infections in immuno-depressed subjects.

The antimycotic activity of the compounds of formula II-A object of thepresent invention has been evaluated also in comparison with the one ofthe corresponding enantiomers.

In some cases the activity of these enantiomers was comparable with theone of the reference compounds, nevertheless it is overall markedlylower than the one of the compounds of formula II-A object of thepresent invention.

The in vivo antimycotic activity has been estimated, bothintraperitoneally and orally, in a Candida model experimentally inducedin mouse against Candida albicans strains sensible to Fluconazole andItraconazole.

The 50% protective dose (PD₅₀) of the compounds of formula II-A has beendetermined in the in vivo tests and it showed to be at least comparableto the one of the reference compounds.

Therefore the compounds of formula II-A object of the present inventionare active on wide range deep mycosis, but particularly againstopportunistic pathogenic agents responsible of infections inimmuno-depressed subjects, they are topically, orally and parenterallyadministrable and endowed with a good therapeutic index.

Then the compounds of formula II-A are useful in the human or veterinarytreatment and prophylaxis of systemic and mucosal infections due tofungi and yeasts.

As just emphasised, the sound pharmacological activity of the compoundsof formula II-A, shown also against strains resistant to antimycoticsused in therapy and against recently isolated strains responsible ofinfections in immuno-depressed subjects, is particularly surprisingconsidering that the compounds disclosed by the European patentapplication No. 315946 were said to be immunising agents for fungalpathologies and phytogrowth regulating agents on useful growings, boththeir activities being limited to the agricultural use.

The pharmacological activity of the compounds of formula II-A is stillmore surprising considering that such compounds, while showing somestructural moieties in common with the azole tertiary alcohols describedby the literature, are in the same time characterized by the presence ofa two carbon atoms chain between the carbon atom bearing the hydroxygroup and the oxygen or sulphur atom, and of a simple polyfluorinatedalkyl in the ether or sulphur containing moiety.

As far as we know, the combination of these structural characteristicshas never been described by the literature in the rank of the compoundsof the azole tertiary alcohols class with antimycotic activity for humanand animal use.

For the human and veterinary use the compounds of formula II-A may beadministered in admixture with a suitable carrier selected in view ofthe administration route. Therefore a further object of the presentinvention comprises pharmaceutical compositions containing atherapeutically effective amount of one of the compounds of formula II-Ain admixture with a pharmaceutically acceptable carrier.

For example, the compounds of formula II-A or the salts thereof may beorally administered as tablets, capsules, solutions or suspensions.

As for the parenteral administration, for example via intravenous,intramuscular or subcutaneous route, the compounds of formula II-A orthe salts thereof are in form of sterile aqueous solution.

Alternatively, the compounds of formula II-A or the salts thereof may beadministered as suppositories or pessaries.

As for the topical administration, the compounds of formula II-A or thesalts thereof are preferably formulated as creams or powders.

As for the oral or parenteral administration the daily dosage of thecompound of formula II-A generally ranges between 0.1 and 50 mg/kg,preferably between 1 and 20 mg/kg, to be subdivided in one or morespaced out doses.

For better illustrating the present invention the following examples areprovided.

EXAMPLE 1 Enzymatic Kynetic Resolution of (±)3-(2,4-difluorophenyl)-2,2-dimethyl-4-(1H-1,2,4-triazol-1-yl)-1,3-butandiol

a) Isomer (+)

The enzyme (3 g; lipase from Candida cilindracea type VII--Sigma) wasadded under vigorous stirring at 30° C. to a solution of (±)3-(2,4-difluorophenyl)-2,2-dimethyl-4-(1H-1,2,4-triazol-1-yl)-1,3-butandiol(2,97 g; 10 mmoles) in chloroform (60 ml) and vinyl acetate (1.9 ml; 20mmoles).

After 72 hours the suspension was filtered, and further enzyme was added(3 g) while maintaining under stirring at 30° C. for further 24 hours.The suspension was filtered, the solid washed with chloroform (20 ml)and the organic phase evaporated under reduced pressure.

The residue was chromatographed on silica gel (eluent ethylacetate:hexane=60:40) to yield (-)3-(2,4-difluorophenyl)-3-hydroxy-2,2-dimethyl-4-(1H-1,2,4-triazol-1-yl)butylacetate (1.63 g; e.e. 88% chiral HPLC--column CHIRACEL-OD, eluenthexane:isopropanol=70:30) and (+)3-(2,4-difluorophenyl)-2,2-dimethyl-4-(1H-1,2,4-triazol-1-yl)-1,3-butandiol(1.3 g; e.e. 79% chiral HPLC).

The isomer (+) thus obtained (1.3 g) was treated again under the sameconditions with the lipase for 17 hours.

After the same treatment and isolation by flash chromatography (+)3-(2,4-difluorophenyl)-2,2-dimethyl-4-(1H-1,2,4-triazol-1-yl)-1,3-butandiol(0.92 g; yield 62%) was obtained as a white solid.

[α]D²⁰ =+43.4° (c=1%, methanol); Chiral HPLC=e.e. 95%; m.p. 116.5-117.5°C.;

¹ H-NMR identical to the one of the racemic mixture.

b) Isomer (-)

(-)3-(2,4-difluorophenyl)-3-hydroxy-2,2-dimethyl-4-(1H-1,2,4-triazol-1-yl)butylacetate obtained in the previous step (a) (1.63 g) was dissolved in a0.05M pH 7 phosphate buffer (400 ml) and acetone (50 ml).

The solution under stirring at 30° C. was added with the enzyme (1.6 g;lipase from Candida cilindracea type VII) and the pH was maintainedduring the subsequent addition of 0.1 M sodium hydroxide.

After 5 hours pH was brought to 4 by concentrated hydrochloric acid andthe extraction was effected with ethyl acetate (3×100 ml).

The organic phase was anhydrified over sodium sulphate and evaporatedunder reduced pressure to yield an oily residue.

After flash chromatography on silica gel (eluent ethylacetate:hexane=60:40) (-)3-(2,4-difluorophenyl)-2,2-dimethyl-4-(1H-1,2,4-triazol-1-yl)-1,3-butandiol(1.20 g; yield 80%) was isolated as a white solid.

[α]D²⁰ =-46.2° (c=1%, methanol); Chiral HPLC=e.e. 98.4%; m.p. 117-118°C.; ¹ H-NMR identical to the one of the racemic mixture.

EXAMPLE 2 Preparation of (-)(2S,3R)-3-(2,4-difluorophenyl)-2-methyl-4-(1H-1,2,4-triazol-1-yl)-1.3-butandiol

A solution of(2R,3R)-3-(2,4-difluorophenyl)-3-hydroxy-2-methyl-4-(1H-1,2,4-triazol-1-yl)butanoicacid (0.88 g; 2.97 mmoles) in anhydrous tetrahydrofuran (6.5 ml) wasdropwise added with boron trifluoroetherate (0.365 ml; 2.97 mmoles) andthe mixture was refluxed for 30 minutes.

The resulting solution was dropwise added at 55° C. with a 10M solutionof boranedimethylsulphide in tetrahydrofuran (0.356 ml; 3.56 mmoles) andthe reflux was maintained for 6 hours.

After cooling to 4° C. a solution water:tetrahydrofuran=1:1 (3 ml) then5M soda (8 ml) were added and the mixture was refluxed for 12 hours.

The tetrahydrofuran was evaporated under reduced pressure, the mixturewas extracted with chloroform (5×10 ml), the organic phase wasanhydrified and evaporated to yield a solid residue which wascrystallised from benzene:hexane=1:1 (20 ml) to give (-)(2S,3R)-3-(2,4-difluorophenyl)-2-methyl-4-(1H-1,2,4-triazol-1-yl)-1,3-butandiol(0.76 g; yield 90%).

m.p. 93-95° C. [α]D²⁰ =-59.6° (c=1%, methanol); ¹ H-NMR (CDCl₃): 7.93(s, 1H); 7.74 (s, 1H); 7.48-7.33 (m, 1H); 6.83-6.68 (m, 2H); 5.40(s-broad, 1H); 4.97 (d, 1H); 4.78 (d, 1H); 4.00 (dd, 1H); 3.80 (dd, 1H);3.10 (s-broad, 1H); 2.45-2.25 (m, 1H); 0.85 (d, 3H).

Similarly operating the following compounds were prepared.

(-)(2S,3R)-3-(2,4-dichlorophenyl)-2-methyl-4-(1H-1,2,4-triazol-1-yl)-1,3-butandiol

yield 87%; m.p. 108-110° C.; [α]D²⁰ =-90.7° (c=1%, methanol).

(+) (2R,3S)-3-(2,4-dichlorophenyl)-2-methyl-4-(1H-1,2,4-triazol-1-yl)-1,3-butandiol

yield 91%; m.p. 110-111° C.; [α]D²⁰ =+87.0° (c=1%, methanol).

(-)(2S,3R)-2-ethyl-3-(2,4-difluorophenyl)-4-(1H-1,2,4-triazol-1-yl)-1,3-butandiol

yield 90%; m.p. 75-77° C.; [α]D²⁰ =-41.1° (c=l%, methanol), Chiral HPLC(hexane:isopropanol=70:30)=e.e. 92%; ¹ H-NMR (CDCl₃): 7.89 and 7.71 (2s,2H, 2CH-Triaz.); 7.40-6.62 (m, 3H, Ar); 5.30 (s-broad, 1H, OH);4.98-4.70 (m, 2H, CH₂ -Triaz.); 4.12-3.87 (m, 2H, CH₂ --OH); 2.80(s-broad, 1H, CH₂ --OH); 2.05-1.93 (m, 2H, *CH--CH₂ --CH₃); 1.60-0.98(m, 2H, CH--*CH₂ --CH₃); 0.81 (t, 3H, CH--CH₂ --*CH₃).

EXAMPLE 3

Preparation of (-)2-(2,4-difluorophenyl)-4-(1,1,2,2-tetrafluoroethoxy)-3,3-dimethyl-1-(1H-1,2,4-triazol-1-yl)-2-butanol(Compound 1A)

A solution of (-)3-(2,4-difluorophenyl)-2,2-dimethyl-4-(1H-1,2,4-triazol-1-yl)-1,3-butandiol(5 g; 16.8 mmoles), prepared as described in Example 1, anddimethylsulphoxide (8 ml) in toluene (60 ml) under stirring at -5° C.was added with potassium hydroxide in dust (533 mg; 9.5 mmoles).

The reaction atmosphere was substituted with tetrafluoroethylene and themixture was maintained under stirring at -5° C. for 90 minutes.

After an addition of water (120 ml), the organic phase was washed with5% hydrochloric acid (80 ml) and treated with anhydrous sodiumhydrocarbonate (6.5 g) under stirring for 30 minutes.

The liquid phase was filtered and the solvent evaporated under reducedpressure to yield an oily residue which was purified by flashchromatography (eluent hexane: ethyl acetate=6:4) to give the compound1A (5.2 g; yield 79%) as a white solid.

[α]D²⁰ =-48.5° (c=1%, methanol); Chiral HPLC=e.e. ≧99.8%; m.p. 46-47°C.; ¹ H-NMR (CDCl₃): 8.01 (d, 1H, CH-Tetr.); 7.69 (s, 1H, CH-Cat);7.63-6.56 (m, 3H, Ar); 5.70 (tt, 1H, JHF=53.4 Hz, CHF₂); 5.33 (s-broad,1H, OH); 5.29-4.41 (m, 2H, CH₂ -Triaz.); system AB: VA=4.19, VB=3.75,JAB=9.8 Hz, CH₂ --O; 1.06 (d, 3H, JHF=2.4 Hz, CH₃); 0.98 (s, 3H, CH₃).

Similarly operating the following compounds were prepared.

(+)2-(2,4-difluorophenyl)-4-(1,1,2,2-tetrafluoroethoxy)-3,3-dimethyl-1,2,4,-triazol-1-yl-)-2-butanol(Compound 1B)

yield 87%; [α]D²⁰ =-56.1° (c=1%, methanol); Chiral HPLC(hexane:isopropanol=90:10)=e.e. 99% nitrate (crystallized from isopropylether/acetonitrile); m.p. 89-91° C.; ¹ H-NMR (CDCl₃): 9.70 (s, 1H,CH-Triaz.); 8.05 (s, 1H, CH-Triaz.); 7.37-6.67 (m, 3H, Ar); 6.77(s-broad, H⁺); 5.80 (tt, 1H, JHF=52.6 Hz, CHF₂); system AB: Va=5.11,Vb=4.87, Jab=14.2 Hz, *CH₂ -Triaz.; part AB of system ABX: Va=4.42,Vb=4.21, Jab=11.1 Hz, Jax=7.7 Hz, Jbx=2.6 Hz; CH₂ --O; 2.41-2.30 (m, 1H,*CH--CH₂); 1.32-1.18 (m, 2H, CH--*CH₂); 0.83 (d, 3H, JHH=7.1 Hz, CH₃).

Example 4

In vitro Antimycotic Activity

The activity inhibiting the growth of mycetes has been evaluated by themacromethods of the scalar brothdilution in geometrical progression (M.R. McGinnis and M. G. Rinaldi, "Antimycotic drugs: mechanisms of action,drug resistance, susceptibility testing and assay of activity inbiological fluids", in Antibiotic in Laboratory Medicine, Ed. V. Lorian,Baltimora 1991).

As coltural medium Yeast Nitrogen Base broth (YNB) and SabouraudDextrose broth (SDB) were employed for yeasts and moulds respectively.

The results obtained in SDB (after incubation at 28° C. for 7 days) areexpressed as minimum inhibiting concentration (MIC) the growing of themycetes, whereas the results obtained in YNB (after incubation at 35° C.for 48 hours) are expressed as concentration inhibiting at 50% (IC₅₀)the growing of the yeast. Fluconazole has been taken as referencecompound.

In the following Tables the data of the in vitro antimycotic activityagainst Candida albicans, Aspergillus fumigatus, Cryptococcus neoformansand Trichophyton mentagrophytes of some representative compounds offormula II are set forth.

TABLE 1

In vitro antimycotic activity of the compounds 1A, 3A, their respectiveenantiomers 1B, 3B, the compound 2A and the reference compoundFluconazole against C. albicans, C. neoformans, T. mentagrophytes and A.fumigatus.

(+)(2S,3R)-2-(2,4-dichlorophenyl)-4-(1,1,2,2-tetrafluoroethoxy)-3-methyl-1-(1H-1,2,4-triazol-1-yl)-2-butanol(Compound 3B)

colourless oil--yield 63%; [α]D²⁰ =+76.5° (c=1%, methanol); Chiral HPLC(hexane:isopropanol=90:10)=e.e. 99.8% nitrate (crystallised fromisopropyl ether); m.p. 120.9-121.9° C.; ¹ H-NMR (CDCl₃): 9.67 and 8.05(2s, 2H, Triaz.); 8.21 (s-broad, 2H, OH e HNO₃); 7.44 (d, 1H, JHH=8.6Hz, C--*CH--CH--C--Cl); 7.35 (d, 1H, JHH=2.2 Hz, C--Cl--CH--C--Cl); 7.09(dd, 1H, C--CH--*CH--C--Cl); 5.82 (tt, 1H, JHF=52.8 Hz, CHF₂); systemAB: Va=5.67, Vb=4.89, Jab=14.3 Hz, *CH₂ -Triaz.; part AB of system ABX:Va=4.44, Vb=4.09, Jab=10.8 Hz, Jax=8.4 Hz, Jbx=4.0 Hz; CH₂ --O;3.32-3.16 (m, 1H, *CH--CH₃); 0.71 (d, 3H, JHH=7.0 Hz, *CH₃ --CH).

(-)(2R,3S)-3-etil-2-(2,4-difluorophenyl)-4-(1,1,2,2-tetrafluoroethoxy)-1-(1H-1,2,4-triazol-1-yl)-2-butanol(Compound 4A)

yield 87%; [α]D²⁰ =-56.1° (c=1%, methanol); Chiral HPLC(hexane:isopropanol=90:10)=e.e. 99% nitrate (crystallised from isopropylether/acetonitrile); m.p. 89-91° C.; ¹ H-NMR (CDCl₃): 9.70 (s, 1H,CH-Triaz.); 8.05 (s, 1H, CH-Triaz.); 7.37-6.67 (m, 3H, Ar); 6.77(s-broad, H⁺); 5.80 (tt, 1H, JHF=52.6 Hz, CHF₂); system AB: Va=5.11,Vb=4.87, Jab=14.2 Hz, *CH₂ -Triaz.; part AB of system ABX: Va=4.42,Vb=4.21, Jab=11.1 Hz, Jax=7.7 Hz, Jbx=2.6 Hz; CH₂ --O; 2.41-2.30 (m, 1H,*CH--CH₂); 1.32-1.18 (m 2H, CH--*CH₂); 0.83 (d, 3H, JHH=7.1 Hz, CH₃).

EXAMPLE 4

Preparation of (-)(2R,3S)-2-(2,4-dichlorophenyl)-4-(1,1,2,2-tetrafluoroethoxy)-3-methyl-1-(1H-1,2,4-triazol-1-yl)-2-butanol(Compound 3A)

Alternatively to the purification procedure of Example 3, the titlecompound as a crude (86.2 g) was dissolved in 173 ml of ethanol at atemperature comprised between 30 and 35° C., then filtered to obtain aperfectly clear solution which was dropped, at 20° C. in 90 minutes,into a mixture of water (260 ml) and ethanol (87 ml).

The mixture was heated to 35° C. kept under stirring for 90 minutes,cooled to 2° C. and again stirred for 60 minutes.

The insoluble solid was filtered, washed with a mixture ethanol/water1:1 and dried in a oven under vacuum at 50° C. till a constant weight,thus yielding 74 g of (-)(2R,3S)-2-(2,4-dichlorophenyl)-4-(1,1,2,2-tetrafluoroethoxy)-3-methyl-1-(1H-1,2,4-triazol-1-yl)-2-butanol(Compound 3A) with a purity >99%;

[α]D²² =-83.6 (c=1%, methanol); Chiral HPLC(hexane:isopropanol=90:10)=e.e. 99,5%; m.p. 79-80° C.; ¹ H-NMR (CDCl₃):7.80 and 8.05 (2s, 2H, Triaz.); 7.42 (d, 1H, JHH=8.6 Hz,C--*CH--CH--C--Cl); 7.27 (d, 1H, JHH=2.1 Hz, C--Cl--CH--C--Cl); 7.05(dd, 1H, C--CH--*CH--C--Cl); 5.75 (tt, 1H, JHF=53.4 Hz, CHF₂); systemAB=VA=5.48, VB=4.54, JAB=14.4 Hz, *CH₂ -Triaz.); 5.09 (broad signal, 1H,OH); 4.54-3.98 (m, 2H, CH₂ --O); 3.14-2.97 (m, 1H, *CH--CH₃); 0.67 (d,3H, JHH=7.1 Hz, *CH₃ --CH).

EXAMPLE 5

Preparation of 3-(2,4-dichlorophenyl)-3-hydroxy-2-methyl-4-(1H,1,2,4-triazol-1-yl)methansulphonic acid butyl ester

A solution of (+)(2R,3S)-3-(2,4-dichlorophenyl)-2-methyl-4-(1H-1,2,4-triazol-1-yl)-1,3-butandiol(2.95 g; 9.31 mmoles), prepared as described in Example 2, in methylenechloride (90 ml) was added, under nitrogen atmosphere, firstly withtriethylamine (1.94 ml; 13.9 mmoles), then with mesyl chloride (0.735ml; 9.46 mmoles) at 0° C.

The reaction was carried out for 20 minutes while maintaining thetemperature at 0° C., then the reaction mixture was poured into water(60 ml).

The phases were separated and the aqueous one was further extractedtwice with methylene chloride.

The collected organic phases were washed with a 5% aqueous solution ofsodium hydrocarbonate, then a saturated aqueous solution of sodiumchloride, and anhydrified over sodium sulphate.

The solvent evaporation yielded 3.64 g (100%) of3-(2,4-dichlorophenyl)-3-hydroxy-2-methyl-4-(1H-1,2,4-triazol-1-yl)methansulphonicacid butyl ester as a white solid which was used as such in the nextstep.

¹ H-NMR (CDCl₃): 7.84 and 7.79 (2s, 2H, H-Triaz.); 7.50-7.00 (m, 3H,H-phenyl); 5.56 and 4.56 (2d, 2H, CH₂ -Triaz.); 4.77 and 4.28 (2dd, 2H,CH₂ -OMs); 3.20-3.10 (m, 1H, CH); 3.10 (s, 3H, CH₃ mesylate); 0.70 (d,3H, CH₃).

EXAMPLE 6

Preparation of thioacetic acidS-[3-(2,4-dichlorophenyl)-3-hydroxy-2-methyl-4-(1H-1,2,4-triazol-1-yl)butyl]ester

3-(2,4-dichlorophenyl)-3-hydroxy-2-methyl-4-(1H-1,2,4-triazol-1-yl)methansulphonicacid butyl ester (9.3 mmoles), prepared as described in Example 5, wasdissolved in ethanol (95 ml) and added with potassium thioacetate (2.12g). The mixture was refluxed for 2 hours, then cooled in ice and theresulting precipitate was filtered and washed with iced methylenechloride. The filtrate was concentrated under reduced pressure, taken upwith water (60 ml) and extracted more times with methylene chloride. Theorganic phases were collected and anhydrified over sodium sulphate, thusyielding 3.5 g of a reddish crude which was purified by flashchromatography on silica gel (eluent ethyl acetate:hexane=1:1) thusyielding 1.72 (49%) of thioacetic acidS-[3-(2,4-dichlorophenyl)-3-hydroxy-2-methyl-4-(1H-1,2,4-triazol-1-yl)butyl]ester.

¹ H-NMR (CDCl₃): 7.85 and 7.76 (2s, 2H, H-Triaz.); 7.50-7.00 (m, 3H,H-phenyl); 5.62 and 4.75 (2d, 2H, CH₂ -Triaz.); 3.60-3.45 and 2.80-2.65(2m, 2H, CH₂ --SCOCH₃); 2.75 (m, 1H, CH); 2.39 (s, 3H, CH₃ thioacetate);0.70 (d, 3H, CH₃).

EXAMPLE 7

Preparation of(2R,3R)-3-(2,4-dichlorophenyl)-3-hydroxy-2-methyl-4-(1H-1,2,4-triazol-1-yl)-1-butanthiol

Under nitrogen atmosphere at room temperature a solution of potassiumhydroxide (0.266 g; 4,74 mmoles) in methanol (40 ml) was dropped into asolution of thioacetic acidS-[3-(2,4-dichlorophenyl)-3-hydroxy-2-methyl-4-(1H-1,2,4-triazol-1-yl)butyl]ester, prepared as described in Example 6 (1.72 g; 4.6 mmoles) inmethanol (40 ml).

After reacting for 10 minutes the reaction was quenched with 5% HCl (4ml) diluted with water (20 ml), and the reaction mixture wasconcentrated under reduced pressure, then further diluted with water andextracted more times with methylene chloride. The anhydrification andevaporation of the collected organic phases yielded 1.6 g of a crudewhich was purified by flash chromatography of silica gel (eluent ethylacetate:hexane=1:1). There was thus obtained 1.2 g (80%) of(2R,3R)-3-(2,4-dichlorophenyl)-3-hydroxy-2-methyl-4-(1H-1,2,4-triazol-1-yl)-1-butanthiolas a light oil.

¹ H-NMR (CDCl₃): 7.85 and 7.79 (2s, 2H, H-Triaz.); 7.50-7.00 (m, 3H,H-phenyl); 5.55 and 4.60 (2d, 2H, CH₂ -Triaz.); 3.10-2.90 and 2.80-2.60(2m, 2H, CH₂ --SH); 2.75 (m, 1H, CH); 1.58 (t, 1H, SH); 0.78 (d, 3H,CH₃).

EXAMPLE 8

Preparation of (-)(2R,3R)-2-(2,4-dichlorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoroethylthio)-1-(1H-1,2,4-triazol-1-yl)-2-butanol(Compound 5)

Starting from(2R,3R)-3-(2,4-dichlorophenyl)-3-hydroxy-2-methyl-4-(1H-1,2,4-triazol-1-yl)-1-butanthiol(1.85 g; 5.57 mmoles), prepared as described in Example 7, and followingthe procedure of Example 3, there was obtained a crude which waspurified by flash chromatography (eluent petroleum ether: ethyl acetate6:4) to give 2, 15 g of (-)(2R,3R)-2-(2,4-dichlorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoroethylthio)-1-(1H-1,2,4-triazol-1-yl)-2-butanol(Compound 5) as a white solid.

[α]D²² =-105.7° (c=1%, methanol); Chiral HPLC(hexane:isopropanol=90:10)=e.e. 99.78%; m.p. 61-63° C. ¹ H-NMR (CDCl₃):7.84 (s, 1H, CH-Triaz.); 7.75 (s, 1H, CH-Triaz.); 7.43 (d, 1H, JHH=8.6Hz, *CH--CH--CCl); 7.265 (d, 1H, JHH=2 Hz, Cl--C--CH--C--Cl); 7.08 (dd,1H, CH--*CH--CCl); 5.83 (tt, 1H, JHF=53.8 Hz, CHF₂); system AB=Va=5.50,Vb=4.57, JAB=14.0 Hz, CH₂ -Triaz.); 5.06 (broad signal, 1H, OH);3.47-3.35 (m, 1H, *CH--CH₃); 3.07-2.90 (m, 2H, CH₂ --S); 0.77 (d, 3H,JHH=6.3 Hz,*CH₃ --CH).

EXAMPLE 9

Preparation of (-)(2R,3R)-2-(2,4-dichlorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoroethyisulphinyl)-1-(1H-1,2,4-triazol-1-yl)-2-butanol(Compound 6)

A solution of (-)(2R,3R)-2-(2,4-dichlorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoroethylthio)-1-(1H-1,2,4-triazol-1-yl)-2-butanol(0.48 g; 1.11 mmole), prepared as described in Example 8, in methanol(2.5 ml) was added with 40% hydrogen peroxide (0.073 ml; 0.89 mmole) inthe presence of a catalytic amount of Na₂ WO₄.2H₂ O and left understirring at room temperature for 3 days, then the reaction mixture wastreated with sodium pyrosulphate (Na₂ S₂ O₅) in water, and basified with10% sodium hydroxide.

The mixture was extracted with ether, dried and concentrated undervacuum to yield 0.41 g of an oil which was purified by flashchromatography (eluent ethyl acetate: petroleum ether 4:6).

The resulting oil (0.35 g) was dissolved in ethanol and treated with 70%nitric acid.

The mixture was concentrated under vacuum and gave a solid which wascrumbled with warm isopropyl ether to yield 0.32 g of (-)(2R,3R)-2-(2,4-dichlorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoroethylsulphinyl)-1-(1H-1,2,4-triazol-1-yl)-2-butanol(Compound 6) as a white solid.

m.p. 153-155° C.; ¹ H-NMR (CDCl₃): 9.56 and 9.51 (2s, 1H, CH-Triaz.);8.13 and 8.10 (2s, 1H, CH-Triaz.); 7.58 (s-broad, H+); 7.50-7.08 (m, 3H,Ar); 6.57-5.98 (m, 1H, CHF₂); 5.66-4.87 (m, 2H, CH₂ -Triaz; 3.57-2.79(m, 3H, *CH--CH₃); 3.07-2.90 (m, 2H, CH₂ --S); 0.77 (d, 3H, JHH=6.3Hz,*CH₃ --CH).

EXAMPLE 10

Preparation of (-)(2R,3R)-2-(2,4-dichlorophenyl)-3-methyl-4-(1,1,2,2-tetrachloroethylsulfonyl)-1-(1H-1,2,4-triazol-1-yl)-2-butanol(Compound 7)

A solution of (-)(2R,3R)-2-(2,4-dichlorophenyl)-3-methyl-4-(1,1,2,2-tetrafluoroethylthio)-1-(1H-1,2,4-triazol-1-yl)-2-butanol(0.5 g; 1.15 mmole), prepared as described in Example 8, in methanol(2.5 ml), in the presence of a catalytic amount of Na₂ WO₄. 2H₂ O andconcentrated HCl (0.19 ml; 2.3 mmoles) was added with 40% hydrogenperoxide (0.9 ml; 10.35 mmoles) and heated to 60° C. for 14 hours, thenleft at room temperature overnight.

The reaction mixture was treated with a solution of sodium pyrosulphate(Na₂ S₂ O₅), basified with 10% sodium hydroxide, and extracted withether, dried and concentrated under vacuum to yield 0.49 g of an oilwhich was purified by flash chromatography (eluent ethyl acetate:petroleum ether 1:1).

The resulting oil (0.45 g) was dissolved in ethanol and treated with 70%nitric acid.

The mixture was concentrated under vacuum and gave 0.42 g of (-)(2R,3R)-2-(2,4-dichloro-phenyl)-3-methyl-4-(1,1,2,2-tetrachloroethylsulfonyl)-1-(1H-1,2,4-triazol-1-yl)-2-butanol(Compound 7) as a white solid.

[α]D²² =-63.4° (c=1%, methanol); Chiral HPLC(hexane:isopropanol=90:10)=e.e. 99.7%; m.p. 192-194° C.

¹ H-NMR (CDCl₃): 8.52 (s, 1H, CH-Triaz.); 7.84 (s, 1H, CH-Triaz.);7.58-7.25 (m, 3H, Ar); 7.19 (dt, 1H, JHF=51 Hz, CHF₂); 6.51-6.30 (broadsignal H+); 5.10-4.74 (m, 2H, *CH₂ -Triaz. -AB system); 4.10-3.39 (m,1H, *CH₂ --*CH₃); 0.84 (2d, 3H, JHH=6.1 Hz, CH₃).

EXAMPLE 11

In vitro Antimycotic Activity

A) The activity inhibiting the growth of mycetes has been evaluated bythe macromethods of the scalar broth dilution in geometrical progression(M. R. McGinnis and M. G. Rinaldi, "Antimycotic drugs: mechanisms ofaction, drug resistance, susceptibility testing and assay of activity inbiological fluids", in Antibiotic in Laboratory Medicine, Ed. V. Lorian,Baltimore 1991).

As growing medium Yeast Nitrogen Base broth (YNB) and Sabouraud Dextrosebroth (SDB) were employed for yeasts and moulds respectively.

The results obtained in SDB (after incubation at 28° C. for 7 days) areexpressed as minimum inhibiting concentration (MIC) the growing of themycetes, whereas the results obtained in YNB (after incubation at 35° C.for 48 hours) are expressed as concentration inhibiting at 50% (IC₅₀)the growing of the yeast.

Fluconazole has been taken as reference compound.

In the following Table the data of the in vitro antimycotic activityagainst Candida albicans, Aspergillus fumigatus, Cryptococcus neoformansand Trichophyton mentagrophytes of some representative compounds offormula II are set forth.

                  TABLE 1a                                                        ______________________________________                                        In vitro antimycotic activity of the compounds 1A, 3A, their                    respective enantiomers 1B, 3B, the compound 2A and the reference             compound Fluconazole against C. albicans, C. neoformans,                      T. mentagrophytes and A. fumigatus.                                                   IC.sub.50  (μg/ml)                                                                          MIC (μg/ml)                                               C. albicans          T. menta-                                         Compound 1040 C. neoformans grophytes A fumigatus                           ______________________________________                                        1A       0.0078    0.0078     0.125  4                                          1B 1 1 32 >128                                                                2A(*) 0.0312 0.0312 0.25 8                                                    3A(*) 0.0078 0.0078 0.0625 2                                                  3B(*) 0.25 1 16 >128                                                          Fluconazole 0.5 2 16 >128                                                   ______________________________________                                         (*)the compounds were tested as nitrates.                                

The data set forth in the table show that the compounds object of thepresent invention have an antimycotic activity markedly higher than theone of the reference compound.

B) The activity inhibiting the growth of yeasts and filamentosous fungihas been evaluated by the method of the dilution in agar. The minimuminhibiting concentration at 50% (MIC₅₀) was measured by incorporatinginto the growing medium decreasing concentrations of the testedcompounds thus yielding scalar concentration on basis 2, from 64 to0.00085 μg/ml.

As growing medium casitone agar buffered at pH=7 (casitone 9 g, glucose20 g, sodium citrate 10 g, leaven extract 5 g, agar 18 g, potassiumbiphosphate 0.54 g, sodium biphosphate 3.34 g, distilled water 1,000 ml)was employed.

For evaluating the activity against the yeasts the growing mediumscontaining the scalar concentrations of the compounds to be tested wereinoculated with 1 μl of a suspension of 5×10⁵ of blastospores/mlobtained from 24-48 hours cultures on Sabourad glucosate agar. Theinoculated growing mediums were incubated at 32° C. and the measurementswere effected after 24, 48 hours, after 72 hours for the slow growingyeasts such as the Cryptococcus neoformans

Fluconazole and Itraconazole have been taken as reference compounds.

In the following Table the data of the in vitro antimycotic activityagainst Candida albicans, Candida tropicalis, Candida krusei, Candidaglabrata, Candida parapsilosis, Candida lutitanice, Candida kefyr,Cryptococcus neoformans, Trichosporon spp, Blastoschizomyces capitatus,Geotrichum spp, Prototecha wicherhamii.

                  TABLE 1b                                                        ______________________________________                                        In vitro antimycotic activity expressed as MIC.sub.50 of the compounds 3A      and 2A, and of the reference compounds Fluconazole and Itraconazole           against the listed yeasts.                                                             MIC.sub.50 (μg/ml)                                                         Com-     Compound                                                     Strains pound 2A 3A Fluconazole Itraconazole                                ______________________________________                                        C. albicans                                                                             0.015    0.015     2       0.015                                      C. tropicalis 0.06 0.03 4 0.03                                                C. krusei 0.12 0.007 16 0.03                                                  C. glabrata 0.12 0.12 16 4                                                    C. parapsilosis 0.007 0.007 2 <0.03                                           C. lusitanie 0.015 0.035 1 0.03                                               C. kefyr 0.017 0.017 1 <0.03                                                  C. neoformans 0.06 0.06 16 0.12                                               Trichosporum spp 0.06 0.12 16 0.12                                            B. capitatus 0.25 0.06 8 0.25                                                 Geotrichum spp 0.5 0.12 16 0.1                                                P. wicherhamii 2 0.5 >64 --                                                 ______________________________________                                    

For evaluating the activity against the mycetes the growing mediumscontaining the scalar concentrations of the compounds to be tested wereinoculated with 1 μl of a suspension of 5×10⁵ of spores/ml (conides orendospores) obtained from duly sporulated coltures. The inoculatedgrowing mediums were incubated at 32° C. and the measurements wereeffected after 1, 2, 3, 5, 7 and 10 days.

Fluconazole, Itraconazole and Amphotericin B have been taken asreference compounds.

In the following Table the data of the in vitro antimycotic activityagainst Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger,Aspergillus terreus, Aspergillus alliaceus, Fusariuim verticilloides,Fusarium proliferatum, Pseudoallescheria, Absidia corymbifera, Mucor,Rhyzomucor pusillus, Acremonium, Trichoderma.

                                      TABLE 1c                                    __________________________________________________________________________    In vitro antimycotic activity expressed as MIC.sub.50 of the compounds 3A     and 2A, and of                                                                 the reference compounds Fluconazole, Itraconazole and Amphotericin B         against the                                                                     listed fungi.                                                                       Com- Com-                                                               Strains pound 2A pound 3A Fluconazole Itraconazole Amphotericin             __________________________________________________________________________                                   B                                              A. fumigatus                                                                          0.5  0.5   >64   0.03  0.25                                             A. flavusalis 2 0.5 >6 0.016 0.25                                             A. niger 0.06 0.03 >64 0.016 0.12                                             A. terreus 0.25 0.12 >64 0.016 0.25                                           A. alliaceus 0.5 0.12 >64 0.016 16                                            F. verticilloides 0. 12 0. 12 >64 >8 2                                        F. proliferatum 0.25 0.25 >64 >8 1                                            Pseudoallescheria 0.06 0.06 >64 >16 3                                         A. corymbifera 0.25 0.06 -- 0.03 0.06                                         Mucor 2 0.5 >64 >16 0.03                                                      R. pusillus 0.25 0.016 >64 0.16 0.06                                          Acremonium 4 4 -- >16 16                                                      Trichoderma 0.25 0.12 -- 16 1                                               __________________________________________________________________________

The data set forth in the tables show that the compounds object of thepresent invention have an overall antimycotic activity markedly higherthan the one of the reference compounds.

EXAMPLE 12

In vivo Antimycotic Activity

Albino Charles River mice (CD1 strain) weighing from 23 to 25 g,normally feed with a standard diet and water ad libitum were employed.

Each animal has been intravenously (for systemic candidiasis,crytpococcosis, aspergillosis), intracranially (for intracranialcryptococcosis) or nasally (for pulmonary aspergillosis) treated with asuspension (0.2 ml containing 2.5×10⁷ cells/ml) of the due microorganismin physiological. Right after the injection and after 4, 24 and 48 hoursthe animals has been orally administered (in 2% Arabic gum)geometrically progressive increasing doses of the compound under test.An infected group has been used as control.

The mortality monitoring of the mice has been prolonged up to 14 days.The medium protective dose (PD₅₀) has been calculated by the probitsanalysis (L. Lison--"Statistica applicata alla biologia sperimentale. Laprogrammazione dell'esperimento e l'analisi dei risultati"--CasaEditrice Ambrosiana, 1961) on the basis of the number of animalssurvived at each concentration.

Fluconazole and Itraconazole were taken as reference compounds.

The following Table set forth the data of in vivo antimycotic activityof some compounds representative of the invention after in vivoadministration.

                                      TABLE 2                                     __________________________________________________________________________    Oral antimycotic effectiveness of the compounds 1A, 3A, their respective      enantiomers 1B, 3B, the compound 2A                                             and the reference compounds Fluconazole and Itraconazole, expressed as      medium protective dose (PD.sub.50) after                                        14 days from the infection.                                                              PD.sub.50 (mg/kg/os) at the 14.sup.th day                                                   Com- Com-                                                                              Com- Com- Com-                                pound pound pound pound pound                                               Experimental disease Organism 1A 1B 2A.sup.(a) 3A.sup.(a) 3B.sup.(a)                                                                 Fluconazole                                                                   Itraconazole         __________________________________________________________________________    Systemic candidiasis                                                                       Candida albicans 1040                                                                       0.64 >2  0.32 0.38 4.57                                                                              0.61   1.74                   Systemic cryptococcosis Cryptococcus neoformans 3443 -- -- 8.84 5.48                                                                 63.16 >50                                                                      Intracranial                                                                 cryptococcosis                                                                Cryptococcus                                                                  neoformans ISM                                                                -- -- 14.59 12.5                                                              -- 22.28 >25                                                                   Pulmonary                                                                    aspergillosis                                                                 Aspergillus                                                                   fumigatus MOL-4                                                               -- -- 9.3 6.9 --                                                              >100 >20                                                                       Systemic                                                                     aspergillosis                                                                 Aspergillus                                                                   fumigatus MOL-4                                                               -- -- 3.5 4.5 --                                                              --  10.2             __________________________________________________________________________

The set forth data demonstrate that the compounds of formula II objectof the present invention are active after oral administration at amedium protective dose lower then the one of Itraconazole and at leastequivalent to the one of Fluconazole.

What we claim is:
 1. A compound of formula ##STR8## wherein R₁ ischloro, fluoro, bromo or trifluoromethyl;R₂ is hydrogen, chloro, fluoro,bromo or trifluoromethyl; Z is CH or N; R₃, R₄ and R₅, which are thesame or different, are hydrogen or C₁ -C₄ alkyl, with the proviso thatR₄ is different from R₅ when R₃ is hydrogen; X is O, S, SO or SO₂ ; R₆is a C₁ -C₅ polyfluoroalkyl group containing at least two fluorine atomsand optionally other halogen atoms selected from the group consisting ofchloro and bromo; and the pharmaceutically acceptable acid saltsthereof.
 2. A compound according to claim 1 wherein R₁ is chloro orfluoro, R₂ is hydrogen, chloro or fluoro, R₃ is methyl or ethyl, R₄ andR₅, which are the same or different, are hydrogen, methyl or ethyl, Z isN and R₆ is a 1,1,2,2-tetrafluoroethyl group.
 3. A compound according toclaim 2 wherein R₁ is chloro or fluoro, R₂ is hydrogen, chloro orfluoro, R₃ is methyl or ethyl, R₄ and R₅, which are the same ordifferent, are hydrogen, methyl or ethyl, Z is N, R₆ is a1,1,2,2-tetrafluoroethyl group and X is O or SO₂.
 4. A process for thepreparation of a compound according to claim 1 by reacting apolyfluoroolefin of formula ##STR9## wherein X₁ and X₂, which are thesame or different, are F, Cl or CF₃ ;or by reacting a polyfluorinatedalcohol of formula

    R.sub.7 --CH.sub.2 --OH                                    (IV)

wherein R₇ is a C₁ -C₄ polyfluoroalkyl group containing at least twofluorine atoms and optionally other halogen atoms selected from thegroup consisting of chloro and bromo; with an optically activeintermediate of formula ##STR10## wherein R₁, R₂, R₃, R₄, R₅ and Z havethe same meanings of claim 1, and X is O or S; or a reactive derivativethereof, optionally followed by an oxidation.